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一种用于胰岛功能和形态分析的玻璃平行灌流载玻片。

A Parallel Perifusion Slide From Glass for the Functional and Morphological Analysis of Pancreatic Islets.

作者信息

Schulze Torben, Mattern Kai, Erfle Per, Brüning Dennis, Scherneck Stephan, Dietzel Andreas, Rustenbeck Ingo

机构信息

Institute of Pharmacology and Toxicology, Technische Universität Braunschweig, Braunschweig, Germany.

Center of Pharmaceutical Engineering (PVZ), Technische Universität Braunschweig, Braunschweig, Germany.

出版信息

Front Bioeng Biotechnol. 2021 Mar 5;9:615639. doi: 10.3389/fbioe.2021.615639. eCollection 2021.

DOI:10.3389/fbioe.2021.615639
PMID:33763408
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7982818/
Abstract

An islet-on-chip system in the form of a completely transparent microscope slide optically accessible from both sides was developed. It is made from laser-structured borosilicate glass and enables the parallel perifusion of five microchannels, each containing one islet precisely immobilized in a pyramidal well. The islets can be in inserted via separate loading windows above each pyramidal well. This design enables a gentle, fast and targeted insertion of the islets and a reliable retention in the well while at the same time permitting a sufficiently fast exchange of the media. In addition to the measurement of the hormone content in the fractionated efflux, parallel live cell imaging of the islet is possible. By programmable movement of the microscopic stage imaging of five wells can be performed. The current chip design ensures sufficient time resolution to characterize typical parameters of stimulus-secretion coupling. This was demonstrated by measuring the reaction of the islets to stimulation by glucose and potassium depolarization. After the perifusion experiment islets can be removed for further analysis. The live-dead assay of the removed islets confirmed that the process of insertion and removal was not detrimental to islet structure and viability. In conclusion, the present islet-on-chip design permits the practical implementation of parallel perifusion experiments on a single and easy to load glass slide. For each immobilized islet the correlation between secretion, signal transduction and morphology is possible. The slide concept allows the scale-up to even higher degrees of parallelization.

摘要

开发了一种呈完全透明显微镜载玻片形式的芯片胰岛系统，可从两侧进行光学观察。它由激光结构化硼硅酸盐玻璃制成，能够对五个微通道进行并行灌流，每个微通道都包含一个精确固定在锥形孔中的胰岛。胰岛可通过每个锥形孔上方的单独加载窗口插入。这种设计能够轻柔、快速且有针对性地插入胰岛，并可靠地将其保留在孔中，同时允许足够快速地更换培养基。除了测量分级流出物中的激素含量外，还可以对胰岛进行并行活细胞成像。通过显微镜载物台的可编程移动，可以对五个孔进行成像。当前的芯片设计确保了足够的时间分辨率来表征刺激 - 分泌偶联的典型参数。这通过测量胰岛对葡萄糖刺激和钾去极化的反应得到了证明。在灌流实验后，胰岛可以取出进行进一步分析。对取出的胰岛进行的活死检测证实，插入和取出过程对胰岛结构和活力无害。总之，当前的芯片胰岛设计允许在单个且易于加载的载玻片上实际进行并行灌流实验。对于每个固定的胰岛，可以研究分泌、信号转导和形态之间的相关性。载玻片概念允许扩展到更高程度的并行化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eef4/7982818/0a3f4ce57206/fbioe-09-615639-g013.jpg
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